Interleukin-1 stimulates prostaglandin E production by human trophoblast cells from first and third trimesters.

Prostaglandins (PGs) play a major role during implantation and labor, and their level is regulated by various cytokines. Interleukin-1 (IL-1) is a known mediator of prostaglandin E (PGE) production in various cell types, including endothelial, amniotic, and endometrial cells; however, its role in the regulation of PGE production in the trophoblast cells is yet unknown. As IL-1 and PGE are both known to be synthesized in the human trophoblast cells, we examined the possibility that IL-1 regulates PG production in human trophoblast cells. To this end, use was made of first and third trimester trophoblast cells, obtained from first trimester terminations of pregnancies and elective cesarean sections. The trophoblast cells were separated by trypsin degradation and fractionation on Percoll gradients, and cultured for 18 h under serum-free conditions in the absence or presence of IL-1 (10 ng/mL). IL-1 induced a 5-fold increase in PGE production, a response that was cell density, time, and dose dependent. IL-1-induced PGE biosynthesis was prevented in the presence of either IL-1 receptor antagonist or the soluble IL-1 receptor, suggesting a receptor-mediated response. Significantly, de novo production of PGE by trophoblast cells in the absence of IL-1 was also markedly (50%) reduced by either the IL-1 receptor antagonist or the soluble IL-1 receptor, further supporting the notion that IL-1 is involved in PGE synthesis even under basal conditions. Transforming growth factor-beta 1, a putative modulator of the effects of IL-1, significantly attenuated IL-1-stimulated PGE production, supporting the possibility that transforming growth factor-beta 1 may serve as a regulator of the effects of IL-1 in trophoblast cells. These observations suggest a pivotal role of IL-1 in the regulation of PGE economy by trophoblast cells. As trophoblast cells are in intimate contact with maternal cells, understanding the regulation of PGE levels may explain crucial processes at the feto-maternal interface, including implantation of the developing blastocyst, immunosurveilance, and the initiation of labor.

Cytokine-mediated regulation of type IV collagenase expression and production in human trophoblast cells.

The invasive property of trophoblast cells is dependent on the activity of proteolytic enzymes of the metallo- and serine proteases family. Interleukin-1 (IL-1) was found to be involved in the regulation of these proteases in various systems, serving as an important modulator in trophoblast physiology (e.g. induction of hCG beta, cytokines, and others). Therefore, consideration is given in this report to the role of IL-1 in the regulation of metalloprotease activity in human trophoblasts. Human trophoblast cells were isolated from first trimester placentas by trypsin degradation and Percoll fractionation. Primary cell cultures of first trimester trophoblasts constitutively elaborated two species of collagenase type IV (92 and 72 kDa), as assessed in gelatin matrix. Treatment with IL-1 further augmented the 92-kDa type IV collagenase secretion in a dose-dependent manner. Furthermore, IL-1 significantly (P < 0.01) increased 92-kDa collagenase gene expression by trophoblast cells, as determined by solution hybridization/ribonuclease protection assay. Both the increase in gene expression and protein biosynthesis of the 92-kDa collagenase type IV were neutralized by the soluble IL-1 receptor, indirectly suggesting a receptor-mediated response. Interestingly, transforming growth factor-beta a putative modulator of IL-1 induced effects, was shown to induce the 92-kDa collagenase type IV secretion as well. These results provide indirect evidence supporting the idea that IL-1 and transforming growth factor-beta may play an intermediary role in trophoblast invasion at the feto-maternal interface by regulating trophoblast expression of 92-kDa type IV collagenase, a protease of prime importance in trophoblast invasion.

Interleukin-1-mediated stimulation of prostaglandin E production is without effect on plasminogen activator activity in human granulosa lutein cell cultures.

In continuation of earlier observations on the involvement of interleukin-1 (IL-1) in ovarian function, we examined the ability of IL-1 to modulate plasminogen activator (PA) activity and prostaglandin (PG) synthesis in human granulosa lutein cells (GLCs). Toward this goal, GLCs were obtained from women undergoing in vitro fertilization, preincubated with 10% fetal calf serum for 48 h, and subsequently cultured for 48 h in serum-free media in the absence or presence of IL-1 beta (10 ng/mL). Cellular PA activity was measured by plasminogen-dependent cleavage of the chromogenic substrate H-D-valyl-L-leucyl-L-lysine-p-nitroanilide (S-2251). Prostaglandin E (PGE) levels were assayed by conventional RIA. Exposure of GLCs to IL-1 resulted in a 50% increase in PGE production, a 33% suppression of PA activity, and a 75% increase in the ability of the corresponding conditioned media to inhibit exogenous urokinase activity. The inhibitory capacity was attributable to an IL-1-mediated increase in PA inhibitor type-1 (PAI-1) production, inasmuch as urokinase inhibition could be abolished by the administration of a polyclonal antihuman PAI-1 immunoglobulin G. IL-1 treatment had no effect on plasmin or trypsin inhibition. Exposure of GLCs to IL-1 receptor antagonist abolished the ability of IL-1 to enhance PA inhibitory activity and PGE production, thereby establishing specific IL-1 receptor-mediated effects. The ability of IL-1 to suppress PA activity and to produce PAI-1 persisted in the presence of indomethacin, a potent inhibitor of PG synthesis. Likewise, transforming growth factor-beta 1 suppressed the ability of IL-1 to stimulate PGE production without affecting the IL-1-induced effects on the PA system. The present findings suggest a pluripotent response of GLCs to IL-1, characterized by the induction of PAI-1 and the suppression of PA occurring concurrent with, but independent of, PG production. These observations support the potential involvement of IL-1 in the regulation of human ovulatory processes.

High levels of human chorionic gonadotropin retard first trimester trophoblast invasion in vitro by decreasing urokinase plasminogen activator and collagenase activities.

Trophoblast cells of the blastocyst and of the first trimester placenta penetrate the endometrial basement membrane during the process of implantation and placental development. However, this invasive capacity seems to be restricted to the fetomaternal interface, as few trophoblast cells can be identified in the decidua, and trophoblasts rarely penetrate the maternal blood vessels. We have shown that the high invasive ability of first trimester human trophoblasts in vitro depends on collagenase activated by plasmin generation. In our study we used invasive first trimester trophoblast cells in conjunction with as in vitro amnion invasion assay to assess the role of hCG in the invasive process. hCG inhibited trophoblast invasion capacity in a dose-dependent fashion but exerted no effect on the ability of the trophoblasts to attach to the basement membrane. The activity of collagenase by trophoblasts (determined by zymography) was down-regulated by hCG, again in a dose-dependent manner. In contrast, hCG had no effect on production of the tissue inhibitor of metalloproteinases. Similar inhibitory effects of hCG on urokinase-plasminogen activator (uPA) and the activity of trophoblast-conditioned media were shown (measured by degradation of S-2444). The hCG effect on collagenase production was not mediated by the expression of procollagenase messenger RNA (mRNA), the expression of the mRNA encoding tissue inhibitor of metalloproteinase, or the expression of uPA mRNA, suggesting posttranscriptional control of hCG action. High levels of hCG attenuated the activity of commercial uPA but had no effect on commercial collagenase activity. These observations suggest that hCG may play a role in the trophoblast invasion process by inhibition of uPA activity, in turn decreasing collagenase activity and thereby reducing trophoblast cell invasion.

Interleukin-1 beta inhibits progesterone accumulation in rat corpora luteal cell cultures in a mechanism dissociated from its effects on nitric oxide and prostaglandin E accumulation.

The objective of this study was to examine the effect of interleukin-1 beta (IL-1) on progesterone (P) biosynthesis and the potential intermediary involvement of prostaglandin (PG) E and nitric oxide (NO) in P accumulation in PMSG/hCG-primed rat corpora luteal (CL) cell cultures. Exposure of primed CL cells to IL-1 (10 ng/ml) for 48 h resulted in a 65-86% reduction (P < 0.01) in P accumulation concurrent with a 2-3.4-fold increase in PGE content, a 70% increase in PGF2 alpha content and a 1.9-3.3-fold increase in nitrite generation. These effects were abolished by the IL-1 receptor antagonist, suggesting specific IL-1 receptor-mediated effects. Indomethacin, a cyclooxygenase inhibitor, abolished PGE and PGF2 alpha production and attenuated the basal (but not IL-1-stimulated) accumulation of P. N(G)-Nitro-L-arginine (NNLA), a competitive inhibitor of nitrite synthesis, slightly reduced basal P accumulation but had no effect on IL-1-induced suppression of P accumulation. NNLA reduced basal PGE accumulation and IL-1-stimulated PGE accumulation (55 and 61%, respectively). Transforming growth factor beta 1 (TGF-beta 1; 10 ng/ml) significantly attenuated the IL-1-stimulated PGE and NO production (61 and 42%, respectively), but did not affect the ability of IL-1 to suppress P accumulation. Thus, NO, PGF2 alpha and PGE are not obligatory intermediaries of IL-1-mediated suppression of P accumulation in rat CL, but are involved in basal P biosynthesis and NO seems to have a regulatory role in the biosynthesis of PGE. The present observations suggest a pleiotropic response of PMSG/hCG-primed CL cells to IL-1, characterized by an independent suppression of P accumulation and a concomitant increase in NO, PGF2 alpha and PGE generation. Since IL-1 attenuates P accumulation, these findings may imply a direct autocrine/paracrine function for IL-1 in the maintenance or the demise of rat CL.

Germ cell tumors of the ovary arising after dermoid cyst resection: a long-term follow-up study.

OBJECTIVE: To study the long-term ovarian neoplastic consequences of resection of a dermoid cyst. METHODS: The study population comprised 99 patients who were operated on for an ovarian dermoid cyst. Follow-up information was obtained for 91 women for a mean period of 5.06 +/- 2.46 years. RESULTS: Of the 99 women, 18 had bilateral dermoid cysts. Multiple dermoid cysts in a single ovary were found in nine of the women with bilateral cysts and in one of the remaining patients. Two patients developed malignant germ cell tumors, and three developed a recurrent dermoid cyst in an ovary from which a dermoid cyst had previously been extracted. Bilateral or multiple ovarian dermoid cysts were present at the initial operation in four (80%) of these patients. CONCLUSIONS: Women with bilateral or multiple dermoid cysts may include a subgroup of patients with a greater tendency to develop future ovarian germ cell neoplasms.

Do spermatozoa secrete motility enhancing factor?

OBJECTIVES: To determine whether medium conditioned with human spermatozoa was capable of enhancing sperm motility and penetration ability. DESIGN: Paired aliquots of washed spermatozoa were allowed to incubate for nine different incubation periods, ranging from 15 to 240 minutes in 37 degrees C in humidified atmosphere with 5% CO2. After this, they were centrifuged at 600 x g for 6 minutes. The conditioned medium was removed from one tube of each pair and replaced with fresh medium. In the other tube of the same pair the sperm pellet was resuspended in the same medium. In a second set of experiments, conditioned medium was removed from tubes containing samples of spermatozoa after different predefined incubation periods. This was used to replace medium that had been removed from sperm cells that had been incubated for 120 minutes. Motility and penetration of zona-free hamster eggs were assessed. RESULTS: Removal of the incubation medium at times between 15 to 240 minutes resulted in sperm that showed a gradual decrease in motility and penetration ability followed by a gradual increase in motility and penetration ability, i.e., an inverted bell-shaped effect. The addition of conditioned medium obtained after different periods of incubation to spermatozoa where medium was removed after 120 minutes of incubation resulted in an increase in sperm motility and penetration ability. The longer the medium was conditioned with spermatozoa the more prominent the effect on sperm motility and penetration ability, with maximal effect observed with medium conditioned for 120 minutes. CONCLUSIONS: Medium conditioned with human spermatozoa enhances sperm motility and penetration ability.

Interleukin-1-mediated regulation of plasminogen activation in pregnant mare serum gonadotropin-primed rat granulosa cells is independent of prostaglandin production.

OBJECTIVES: This study examines the effects of interleukin-1 (IL-1) on plasminogen activator (PA) activity and prostaglandin (PG) E production in pregnant mare serum gonadotropin (PMSG)-primed granulosa cells and the potential involvement of PGE in the regulation of ovarian plasminogen activation. METHODS: Granulosa cells were obtained from PMSG-primed rat (27-day-old) ovaries and cultured in serum-free conditions for 48 hours in the absence or presence of IL-1 beta (10 ng/mL) with and without transforming growth factor-beta 1 (10 ng/mL). Cellular PA activity was measured through the conversion of plasminogen to plasmin and assay of the plasmin-mediated cleavage of [14C]-labeled globin to acid-soluble products. RESULTS: Exposure of PMSG-primed granulosa cells to IL-1 resulted in a 30% reduction (P < .05) in PA activity. Addition of hCG (1 IU/mL) to the granulosa cell cultures resulted in a 2.3-fold increase in PA activity, an effect significantly attenuated by co-administration of IL-1. The IL-1-mediated inhibition occurred concurrent with a 6.6-fold increase in the ability of the corresponding conditioned media to inhibit exogenous urokinase activity. This latter inhibitory capacity was the result of a significant increase in plasminogen activator inhibitor type 1 (PAI-1), given its abolition by a polyclonal anti-rat PAI-1 immunoglobulin G. The IL-1-mediated effects on PA/PAI-1 were accompanied by a sevenfold increase in PGE content of the spent culture medium. This response was dose dependent. The IL-1 effects on plasminogen activation and PG production were abolished by the IL-1 receptor antagonist, suggesting specific IL-1 receptor-mediated responses. Indomethacin, an inhibitor of PG biosynthesis, prevented the IL-1-induced increase in PGE accumulation but failed to affect the response of the PA system. Transforming growth factor-beta 1, a known regulator of IL-1 action, significantly attenuated the IL-1-stimulated PGE production but did not interfere with the ability of IL-1 to affect the PA system. CONCLUSIONS: The present observations suggest a pleiotropic response of PMSG-primed granulosa cells to IL-1, characterized by the induction of PAI-1 concurrent with but independent of PG production. These findings corroborate and extend earlier observations suggesting that IL-1 affects PA activity and PGE production in immature rat ovaries. Moreover, these observations support our contention that IL-1 may play a major regulatory role in the cellular events leading to ovulation and early corpus luteum formation.

Successful first vaginal birth after cesarean section: a predictor of reduced risk for uterine rupture in subsequent deliveries.

BACKGROUND: Uterine rupture is a catastrophic obstetric complication, most often associated with a preexisting cesarean section scar. Although a vaginal birth after a cesarean is considered safe in modern obstetrics, it is not known whether repeated VBACs increase the risk of rupture, or whether the first VBAC proves the strength and durability of the scar, predicting further successful and less risky vaginal deliveries. OBJECTIVES: To evaluate the effect of repeated vaginal deliveries on the risk of uterine rupture in women who have previously delivered by cesarean section. METHODS: In this retrospective study, 26 VBAC deliveries complicated by uterine rupture were matched for age, parity, and gravidity with 66 controls who achieved VBAC without rupture. The histories, demography, pregnancy, labor and delivery records, as well as neonatal outcome were compared. RESULTS: We found that the risk of rupture decreases dramatically in subsequent VBACs. Of the 40 cases of uterine rupture recorded during the 18 year study period, 26 occurred during VBAC deliveries. Of these, 21 were complicated first VBACs. We also found that the use of prostaglandin-estradiol, instrumental deliveries, and oxytocin had been used significantly more often during deliveries complicated with rupture than in VBAC controls. CONCLUSIONS: Once a woman has achieved VBAC the risk of rupture falls dramatically. The use of oxytocin, PGE2 and instrumental deliveries are additional risk factors for rupture, therefore caution should be exerted regarding their application in the presence of a uterine scar, particularly in the first vaginal birth after cesarean.

Fetal echocardiography: the identification of two of the pulmonary veins from the four-chamber view during the second trimester of pregnancy.

Identification of two of the pulmonary veins as they enter the left atrium is important to the diagnosis of the visceroatrial situs for fetuses with normal and abnormal echocardiographic findings. In this study, we investigated the ability to demonstrate these veins in 734 fetuses varying in gestational age from 13 to 21 weeks. Transvaginal and transabdominal B-mode ultrasound and color flow Doppler were used for the examination. With the use of the transvaginal route, the highest visualization rate, 85-95%, was achieved in fetuses of 14-15 weeks of gestation. With the use of the transabdominal route, the highest visualization rate, 95-97%, was achieved in fetuses of 20-21 weeks of gestation. This examination enabled the correct diagnosis to be made of complex cardiac anomaly.Visualization of the pulmonary veins with B-mode ultrasound aided by color flow Doppler can be accomplished in the majority of fetuses, while the four-chamber view is studied.

Does the menstrual cycle influence cystometry?

Alterations in the hormonal milieu associated with the menstrual cycle appear to influence the dynamic interaction between the bladder and urethra as well as detrusor function, probably because of the common embryological origin of the lower genital and urinary tracts. In this retrospective study we investigated the effect of the menstrual cycle on cystometric diagnosis. A retrospective case note review of 687 consecutive patients attending the urogynecology unit of St Georges University Hospital, a tertiary referral center, was carried out. The study group comprised 57 women with regular menstrual periods. In both patients whose symptoms were adversely affected premenstrually and those whose symptoms were not influenced by the menstrual cycle, the majority of normal cystometric diagnoses were made in the luteal phase: 45.5% vs. 25% (P < or = 0.002) and 38.5% vs. 4.8% (P < or = 0.05), respectively. Diagnoses of genuine stress incontinence, detrusor instability and mixed genuine stress incontinence and detrusor instability were most frequently made in the follicular phase of the cycle. More normal cystometric diagnoses were made in the influenced group (36.8%) than in the uninfluenced group (21%) (P < 0.02). The results of this preliminary study indicate that the timing of cystometric evaluation may influence the detection of a positive diagnosis. The luteal phase may not be the correct time to make an accurate diagnosis, especially in patients whose symptoms are influenced by their menstrual cycle.

Growth hormone status in patients with maturation arrest of spermatogenesis.

The growth hormone levels of 11 azoospermic patients with histological evidence of maturation arrest of unknown aetiology were studied using the clonidine test. The results were compared with the results from clonidine tests on 10 healthy fertile men and on a group of patients with severe oligozoospermia of various aetiologies. The results demonstrated that 10 patients (91%) in the azoospermic group had non-reactive, pathological responses while none of the control group had a pathological response. Of the severe oligozoospermic group, 82% had normal reactive clonidine tests (P < 0.005). We suggest that growth hormone deficiency may be associated with maturation arrest, and the possibility of treatment with growth hormone should be considered.

The prognosis of hypertensive disease of pregnancy accompanied by intrauterine growth retardation in relation to parity.

Twenty-two primiparous women with hypertensive disease of pregnancy (HDP) associated with intra uterine growth retardation (IUGR) were compared with 20 parous women in whom HDP associated with IUGR appeared for the first time in a second or a later pregnancy. Both groups of women were followed up for 10-13 years. The course of the disease among the parous women was more severe as compared to the primiparous women; the mean gestational age at the first increase in blood pressure and gestational age at delivery were significantly earlier in the parous group (33.3 weeks +/- 3.5 v.s. 35.3 +/- 3.2 weeks, p < 0.01, 36.2 weeks +/- 2.2 v.s. 37.6 +/- 1.9 p < 0.01 accordingly). Maternal indication for induction of labor because of uncontrollable hypertension was present in 77% of the cases in the parous group as compared to 31% in the primiparous group (p < 0.05). These complications were present in 66% of subsequent pregnancies in the parous group as compared with 31% in the primiparous group (p < 0.05). Chronic hypertension developed in 33% of the parous group as compared with 20% in the primiparous group. These differences show that the manifestation of HDP is more severe in women in whom HDP with IUGR occur for the first time in a second or a later pregnancy than in those in whom this complication occurs for the first time in a second or a later pregnancy than in those in whom this complication occurs for the first time in their first pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)

Ultrasound-guided transvaginal aspiration in the management of actinomyces pelvic abscess.

BACKGROUND: Increasing reports of intrauterine device (IUD)-related abdominopelvic actinomycosis have been described recently. Surgical therapy has been the usual treatment when tubo-ovarian abscess is identified. CASE: A 38-year-old woman suffering from Actinomyces pelvic abscess unresponsive to medical treatment underwent transvaginal ultrasound-guided needle aspiration. It resulted in marked improvement and avoided surgical treatment. CONCLUSION: Transvaginal needle aspiration of Actinomyces pelvic abscess may be an alternative to surgical therapy, thereby allowing the preservation of pelvic organs.

High concentration of soluble interleukin-2 receptors in ejaculate with low sperm motility.

The various cytokines are involved in infection and immunological reactions and thus may be involved in impairment of sperm function. The present study determined the concentrations of interleukin-1, interleukin-6 and soluble interleukin-2 receptors in ejaculates of normal donors and patients suffering from pure asthenozoospermia and patients suffering from oligoteratoasthenozoospermia. Ejaculates of patients attending the fertility clinic in Hadassah Mount Scopus Hospital, Israel, were evaluated. The patients were divided into three groups: (i) controls (n = 20), (ii) pure asthenozoospermia (n = 30), (iii) oligoteratoasthenozoospermia (n = 36). Significantly higher concentrations of soluble interleukin-2 receptors were found in the ejaculates of patients with pure asthenozoospermia 2243.1 +/- 473.6 mu/ml (mean +/- SE) as compared with controls 673.5 +/- 281.3 mu/ml (P < 0.05) and oligoteratoasthenozoospermic patients 1012.3 +/- 206.9 mu/ml (P < 0.05). Interleukin-1 and interleukin-6 concentrations were comparable in all groups studied. Our results show that human ejaculates contain interleukin-1, interleukin-6 and soluble interleukin-2 receptors. High concentrations of soluble interleukin-2 receptors in the ejaculates of asthenozoospermic patients may be related to either infectious or immunological processes.

Developmental regulation of the expression of 72 and 92 kd type IV collagenases in human trophoblasts: a possible mechanism for control of trophoblast invasion.

OBJECTIVE: During early pregnancy fetal cytotrophoblast cells invade the uterus and penetrate the basement membrane, a property that is characteristic of malignant cells. However, unlike tumor invasion, trophoblast invasion of the uterus is under strict control. This control limits invasion, so that it primarily remains confined to the endometrial aspect of the myometrium and continues only until midgestation. The invasive properties of the trophoblast cells are made possible by the activity of proteolytic enzymes that belong to the metalloproteinases and serine proteinases. Type IV collagenase (metalloproteinase) is considered crucial in the extracellular matrix remodeling that takes place during the invasion process. In this study we set out to characterize the invasive properties of trophoblast cells at different stages of pregnancy. STUDY DESIGN: Human trophoblast cells were isolated from first- and third-trimester placentas by trypsin digestion and Percoll fractionation and were then cultured under serum-free conditions. The invasive ability of trophoblast cells was determined by the in vitro invasion assay, in which the ability of cells to penetrate an artificial basement membrane was examined. Metalloproteinase activity was measured by zymography, and the expression of messenger ribonucleic acid transcripts of 72 and 92 kd type IV collagenases was examined by reverse transcriptase polymerase chain reaction. RESULTS: First-trimester trophoblasts were 3.5 time more invasive in vitro than were third-trimester trophoblast cells (p < 0.005). Although first-trimester trophoblasts secreted both species of type IV collagenase, 72 and 92 kd, in large amounts, third-trimester cells secreted the 92 kd and only minimal amounts of 72 kd type IV collagenase. Moreover, first-trimester trophoblasts secreted significantly more (p < 0.05) 92 kd type IV collagenase than did third-trimester trophoblast. The messenger ribonucleic acid transcript expression of 72 and 92 kd type IV collagenases correlated with the activity of these enzymes secreted by first- and third-trimester trophoblasts. CONCLUSION: The described high in situ invasive capacity of first-trimester trophoblast might be explained by the increased expression and production of 72 kd type IV collagenase and the higher expression of 92 kd type IV collagenase by first-trimester trophoblast cells.

Trophoblasts circulating in maternal blood as candidates for prenatal genetic evaluation.

Trophoblasts have been detected in uterine venous blood, lung parenchyma and maternal blood in the first trimester. Their dilution within maternal leukocytes has been recently estimated to be 1:10(-6). The objectives of this study were to enrich peripheral maternal blood preparations for trophoblast cells, to isolate trophoblasts from the enriched preparation by highly specific markers and to assess fetal cell total number of chromosomal copies by fluorescence in-situ hybridization (FISH). Negative and positive selections for trophoblasts were performed. To assess the efficacy of the enrichment methods, a model mimicking the in-vivo conditions was established. Purified first trimester trophoblasts were prepared from first trimester placentas and were mixed with leukocytes from non-pregnant women in various concentrations. Magnetic beads coupled with antibodies to the common leukocyte antigen (CD45) or to antitrophoblast specific antigens (Trop1, Trop2 and GB25), were attached to peripheral maternal blood cells or to the prepared mixed cell populations. The expression of alpha human chorionic gonadotrophin (alpha HCG) or of human placental lactogen (HPL) by the remaining cells was examined by two means: (i) immunocytochemistry, using monoclonal antibodies against HPL and alpha HCG to stain fetal cells; (ii) reverse transcriptase polymerase chain reaction (RT-PCR), using specific primers for exons of the HPL or alpha HCG mRNAs. Results revealed that HPL- and alpha HCG-expressing cells could be identified in maternal blood only in very rare instances. On the other hand, expression of alpha HCG and HPL by only 100 purified first trimester trophoblasts artificially mixed with peripheral leukocytes at a ratio of 1:10(-5) could be identified by both immunocytochemistry and RT-PCR.(ABSTRACT TRUNCATED AT 250 WORDS)

Successful conservative management of primary nonmetastatic cervical choriocarcinoma.

Primary cervical choriocarcinoma is a rare disease; since 1915 only about 60 cases have been published. The case presented here can be defined as primary cervical choriocarcinoma since it fulfills all the criteria delineated previously.

Expression of gelatinase B by trophoblast cells: down-regulation by progesterone.

OBJECTIVE: It is now accepted that gelatinase B (92 kd type IV collagenase) is involved in blastocyst implantation and trophoblast invasion. However, little is known about the regulation of this enzyme at the fetomaternal interface. Progesterone has been demonstrated to inhibit gelatinase B secretion from endometrial cells, myometrium, and cervical fibroblasts. Interestingly, the promotor of gelatinase B contains a progesterone-responsive element that may explain transcriptional activation of this metalloproteinase by progesterone. It may be hypothesized that progesterone secreted from trophoblast cells, representing the fetal part of the fetomaternal interface, may have a role in the regulation of gelatinase secretion and blastocyst implantation. STUDY DESIGN: To this end, use was made of first-trimester trophoblast cells obtained from first-trimester pregnancy terminations. The trophoblast cells were separated by trypsin degradation and fractionation on Percoll gradients. Metalloproteinase activity was measured by zymography, and the expression of the gelatinase B messenger ribonucleic acid was determined by the solution hybridization/ribonuclease protection assay. RESULTS: Primary cell cultures of trophoblasts from first trimesters of pregnancy constitutively elaborated two species of type IV collagenases (gelatinase A and B) as assessed on a gelatin matrix. Treatment with progesterone decreased the accumulation of a gelatinase B species in a dose-dependent fashion. Administration of a progesterone receptor antagonist onapristone (ZK-98.299) neutralized the progesterone inhibitory effect on the gelatinase B in a dose-dependent fashion, thus supporting the presumption that the progesterone effect is receptor mediated. Progesterone significantly attenuated the expression of gelatinase B by trophoblast cells, an effect that was neutralized by ZK-98.299. CONCLUSION: These observations provide strong indirect support for the participation of progesterone in the regulation of gelatinase B in trophoblast cells. It may be an important regulator of gelatinase production at the fetomaternal interface.